BACKGROUND: Bovine tuberculosis caused by Mycobacterium bovis is an important disease that has negative effects on public health and entails economic loss. Traditional controlling programs for cattle  focus on test and slaughter strategy, and false positive is one of the disadvantages associated with tuberculin skin test. To overcome this limitation, proteins with high specificity have to be utilized. OBJECTIVES: The objective of this study was to clone and express virulent protein CFP-10 from Mycobacterium bovis AN5. METHODS: Full-length genes of cfp-10 were amplified by PCR technique. In parallel, pET23a(+) and PCR products were double digested by EcoRI and HindIII. Ligation was performed at 16˚C followed by transformation into competence E. coli DH5α. After being identified with sequencing, the cloned vector was transformed into E. coli BL21. Induction was performed by isopropyl-β-D-thiogalactopyranoside (IPTG). Urea 8M was used to dissolve the expressed protein in the inclusion body form. Recombinant protein was purified by Nickle-Resin, and urea was eliminated by decreasing the gradient. RESULTS: The CFP-10 gene clone was proved by sequencing method. The CFP-10, as a 10 KDa protein, was confirmed by Western blotting using monoclonal antibodies. Based on the results, the recombinant protein was successfully cloned and expressed. CONCLUSIONS: The results showed that cfp-10 gene was successfully cloned and expressed in prokaryotic system, indicating that this recombinant protein could be utilized in diagnostic kits against bovine tuberculosis in the future.

In bovine tuberculosis caused by Mycobacterium bovis, cattle are known as the main host, and sheep are relatively resistant. Sheep become infected only when there is very close contact between cattle and sheep or when the mycobacterium is abundant in the environment. In the present study, a case of generalized tuberculosis was reported in the sheep of the Sanandaj slaughterhouse.The examined animal carcass was transferred from the slaughterhouse to the University of Kurdistan microbiology laboratory and was dissected under complete health safety conditions. In the carcass examination, necropsy lesions (generalized tubercular granulomas) were visible in different organs and mediastinal lymph nodes, bronchioles, mesentery, and liver portal system. These lesions were seen in the lymph nodes of different body parts, including the lung, liver, digestive system, reproductive system, bladder, and different muscles, including the peritoneal muscle, diaphragm muscle, and abdominal muscles.Diagnostic tests included clinical observations, Ziehl-Neelsen staining for Mycobacterium bacteria, and polymerase chain reaction (PCR) molecular technique. In Ziehl-Neelsen staining, Mycobacterium acid-fast bacilli were seen in red color on the pale blue background of the slide. Also, the product of 499 base pairs was amplified in the specific PCR reaction, which confirmed Mycobacterium bovis.The present report shows that Mycobacterium bovis strains present in the region can be transmitted to sheep and cause disease with severe clinical symptoms. This report is critical, particularly for mixed cattle and sheep farming. Since cattle and sheep are kept together in many breeding farms in Kurdistan province, special attention was paid to the possibility of transmission of bovine tuberculosis to sheep and humans.

A cross-sectional study to determine individual animal prevalence of bovine tuberculosis (BTB) was conducted on 1171 dairy cattle in 12 randomly selected dairy farms in Ethiopia between January 1999 and May 2001 using comparative intradermal tuberculin (CIT) test and bacteriologic study through milk culturing. An overall individual animal prevalence of 46.8% (548 of 1171 animals) and a herd prevalence of 91.7% (11 of 12 farms) were recorded in 12 dairy farms by the CIT test. There were significant (P <0.0001) differences in individual prevalence between farms and breeds (pure Holstein and their crosses with Zebu). There was positive correlation (r = 0.41) between herd size and prevalence of bovine tuberculosis. Furthermore, a negative linear association (R2 = 0.24) was found between mean score of management of the farm and prevalence, indicating that the prevalence of bovine tuberculosis could be improved by sanitary measures. Breed and management affected the prevalence of BTB (R2 = 0.30) as confounding variables. Mycobacterium bovis was isolated from the milk of 13.3% (4 of 30) reactor cows. The widespread occurrence of BTB in the study farms and isolation of M. bovis from the milk of reactor cows signify its economic importance and potential risk to public health. Generalization and improved use of milk pasteurization within all dairy subsectors is recommended, and this would affect the competitiveness of the dairy sector in Ethiopia.

A cross-sectional study was conducted between December 2001 and April 2002 on 94 households and 763 (188 indigenous and 575 crossbred) cattle to determine the prevalence of bovine tuberculosis (BTB) and assess its public health implications in smallholder farms in Wuchale-Jida District, Central Ethiopia. Cluster sampling, single intradermal tuberculin (SIDT) and comparative intradermal tuberculin (CIDT) tests, a questionnaire, and mycobacteriology were used. Based on the CIDT test, herd and individual animal prevalences of BTB were 42.6% and 7.9%, respectively. The individual animal prevalence was significantly affected by herd size (P<0.01), age (P<0.0001) and body condition (P<0.05). Among the interviewed households, 24.5% (23 of 94) had experienced at least one human tuberculosis case in the family. Of these families, 43.5% (10 of 23) had reactor cattle. Nevertheless, no statistically significant association (P>0.05) was observed between reactor cattle and human tuberculosis cases in households. The habit of milk and meat consumption was affected by occupation (P<0.0001) and location of household residence (P<0.001). Although the level of education influenced the habit of milk consumption (P<0.05), it did not impact the habit of meat consumption (P>0.05). Less than half (38.3%; 36 of 94) of the respondents knew about BTB, and only 30.8% (29/94) of the respondents were conscious of its transmission from cattle to humans. Secondary data analysis from Muka-Turri clinic indicated that 85.6% of the human tuberculosis cases were from rural parts of the district. Although the BTB prevalence seems low, its potential risk to public health was important based on food consumption, poor sanitary measures, and the lack of understanding about its zoonosis.

The study highlights the transboundary nature of tuberculosis (TB) in alpacas and the failure of current ante-mortem testing protocols (the tuberculin skin and Enferplex Camelid TB tests) to identify TB-free alpaca herds and individuals for export. Our research and the available literature indicate that the alpaca (Vicugna pacos) is extremely susceptible to Mycobacterium bovis infection, and that testing periodicity fails to take into account that animals do not manifest disease symptoms for a long time. The skin test failed to identify Mycobacterium bovis infection in two alpacas prior to their movement from the UK to Poland. The animals were purchased by a breeding centre in Poland, and were then shown at an international animal exhibition. The last owner of the alpacas before their deaths from TB bought the infected animals unwittingly in order to run rehabilitation activities with disabled children on his farm.

Bovine tuberculosis is one of the most dangerous zoonotic diseases. Despite the near-complete elimination of the disease from cattle breeding in Poland achieved in 2009, its re-emergence is now observed. Globally, the number of human cases is underestimated and the importance of free-living animals as reservoirs of tuberculosis is growing. As a species highly susceptible to Mycobacterium tuberculosis complex infection, the European bison (Bison bonasus) has a role in the transmission of the disease in Poland. The purpose of the investigation was to assess the epidemiological situation of tuberculosis in Polish European bison serologically.

Mycobacterium bovis is the causative agent of bovine tuberculosis, a disease that affects dairy herds throughout the Brazilian territory, constituting a neglected zoonosis transmitted by raw milk and its derivatives. In this study, we evaluated the presence of M. bovis and other mycobacteria in Minas cheese obtained from open fairs in the city of São Paulo between 2012 and 2013. Samples (n = 133) were decontaminated using hexa-cetylpyridinium chloride and seeded on Stonebrink–Leslie medium. The isolates were submitted to molecular identification by TB Multiplex PCR targeting the 16S rRNA gene and amplicon nucleotide sequencing. From 16 cheese samples (12%), we obtained 26 putative colonies of Mycobacterium spp., none of which belonged to any of the Mycobacterium tuberculosis, Mycobacterium avium, or Mycobacterium intracellulare complexes. Phylogenetic analysis showed that sample sequences were grouped in a clade that includes only non-tuberculous mycobacteria with proximity to sequences obtained from Mycobacterium novocastrense (3 sequences), Mycobacterium holsaticum (1 sequence), andMycobacterium elephantis (2 sequences). Although no epidemiological evidence was found regarding the importance of oral transmission of mycobacteria in healthy people, their importance in the immunosuppressed population remains uncertain.

Since 2009, Poland has been recognised as a country officially free of bovine tuberculosis (bTB), although in each year of the last five there were from 8 to 18 outbreaks of the disease. In 2008–2016, the largest number of cattle infected with bovine mycobacteria were eliminated in the Masovian Province (the central region of Poland) and the largest number of outbreaks of this zoonosis were recorded in this area. The close proximity of farms where bTB was found led to the suspicion that tuberculosis could have been transmitted between the affected herds. The aim of the study was the molecular characterisation of the pertinent M. bovis/caprae strains and determination of the epidemiological relationship of various bTB outbreaks. The material for microbiological tests came from 119 cattle (Bos taurus) from nine herds located in five provinces, neighbouring the Masovian Province. Laboratory tests of tissue material gave results confirming tuberculosis in 54 (45%) animals. All strains belonged to the Mycobacterium bovis species. A two-step analysis of genetic affinity allowed 50 strains to be identified as phylogenetically closely related and separated between three genetic clusters consisting of 2 to 27 strains. Based on the results of genotyping, bTB outbreaks were found in three herds, and three transmission chains were identified among these herds.

Introduction: Tuberculosis is a highly infectious disease affecting humans and animals. It is caused by the Mycobacterium tuberculosis complex (MTBC) – Mycobacterium bovis and Mycobacterium caprae, which are aetiological factors of bovine tuberculosis (bTB). In Poland, the bTB eradication programme exists. Animals diagnosed with tuberculosis are in the majority of cases not treated, but removed from their herd and then sanitary slaughtered. Material and Methods: In total, 134 MTBC strains isolated from cattle in Poland were subjected to microbiological analysis. The resistance phenotype was tested for first-line antimycobacterial drugs used in tuberculosis treatment in humans: streptomycin, isoniazid, rifampicin, ethambutol, and pyrazinamide. The strains were isolated from tissues collected post mortem, so the test for drug resistance fulfilled only epidemiological criterion. Results: The analysis of drug-resistance of MTBC strains revealed that strains classified as M. bovis were susceptible to 4 antimycobacterial drugs: isoniazid, rifampicin, streptomycin, and ethambutol, and resistant to pyrazynamide. The strains classified as M. caprae were sensitive to all tested drugs. Conclusion: The results indicate that despite enormously dynamic changes in mycobacterial phenotype, Polish strains of MTBC isolated from cattle have not acquired environmental resistance. The strains classified as M. bovis are characterised by natural resistance to pyrazinamide, which is typical for this species.

All effects taken together, bovine tuberculosis (bTB) has a long-term detrimental effect on bovine herds and many wildlife species in South Africa. The disease is not only found in domestic cattle but also in African buffaloes and has to date been diagnosed in 21 wildlife species, including several rare and endangered species, thus having a potentially serious effect on conservation and biodiversity. In cattle, bTB is mostly characterised by sporadic outbreaks, but bovine herds chronically infected with the clinical disease are not uncommon. Presently, the recognised bTB control strategy in South Africa is based on 'test and slaughter', using the intradermal tuberculin test, followed by the slaughter of animals that have tested positive. Affected herds are placed under veterinary quarantine with movement restrictions until the outbreak is eradicated; this can take several years or last indefinitely if the outbreak cannot be eradicated. The same measures apply to infected buffalo populations, often with no prospect of ever being eradicated. This strategy is neither practical nor viable in the context of a communal farming system and becomes unethical when dealing with valuable wildlife reservoir hosts. Transmission of bTB between wildlife and cattle has been demonstrated and emphasises the need for an effective, affordable and culturally acceptable control strategy to curb the spread of bTB in South Africa. In countries with similar challenges, vaccination has been used and found to be promising for treating wild and domestic reservoir species and may hence be of value as a complementary tool for bTB control in South Africa.